Effect of coding regions on chloroplast gene expression in Chlamydomonas reinhardtii
Effect of coding regions on chloroplast gene expression in Chlamydomonas reinhardtii
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DOI：
10.1263/jbb.95.276
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发表时间：
2003-03-01
影响因子：
2.8
通讯作者：
Shinmyo, A
中科院分区：
文献类型：
--
作者：
Kasai, S;Yoshimura, S;Shinmyo, A
外源基因位于莱茵衣藻叶绿体基因rbcL或psbA的5 '上游区域和rbcL基因的3'末端之间，与莱茵衣藻叶绿体中的内源基因不具有相同的表达模式。本文以莱茵衣藻为材料，选择rbcL、psbA、psbD和atpA四个叶绿体基因，研究叶绿体基因编码区对基因表达的影响。我们构建了由叶绿体基因的启动子、5 '-和3'-非翻译区、不同长度的蛋白质编码区和作为报告基因的细菌β-葡萄糖醛酸酶（GUS）基因（uidA）组成的嵌合基因，并将其导入叶绿体基因组。与先前产生的转化体RG和PG（其中uidA仅融合到每个基因的启动子和5 '-UTR）相比，含有融合到每个基因的编码区的rbcL-uidA和psbA-uidA嵌合基因的转化体显示出uidA mRNA的高表达。转化体中uidA转录物积累的差异是不同转录速率的结果。这一结果表明编码区是rbcL和psbA充分表达所必需的。另一方面，psbD和atpA编码区部分不影响嵌合基因表达。
An exogenous gene, placed between the 5'-upstream regions of the Chlamydomonas reinhardtii chloroplast genes, rbcL or psbA, and the 3'-end of the rbcL gene, do not have the same expression pattern as endogenous genes in the C: reinhardtii chloroplast. Here, we chose four chloroplast genes, rbcL, psbA, psbD and atpA, and examine the effects of chloroplast gene coding regions on gene expression in C, reinhardtii. We constructed chimeric genes composed of the promoter, 5'-and 3'-untranslated regions, varying lengths of protein coding regions of the chloroplast genes, and the bacterial beta-glucuronidase (GUS) gene (uidA) as a reporter gene, and introduced into chloroplast genomes. The transformants, which contained the rbcL-uidA and psbA-uidA chimeric genes fused to the coding region of each gene, showed high expression of uidA mRNA as compared with the previously generated transformants, RG and PG, in which uidA was only fused to the promoter and 5'-UTR of each gene. The difference in the accumulation of uidA transcripts among the transformants was the result of different rates of transcription. This result indicates that the coding region is necessary for sufficient expression of rbcL and psbA. On the other hand, the psbD and atpA coding region portions did not affect chimeric gene expression.